Fluorous peptide microarray, detection process using a fluorous peptide microarray, and process of forming a fluorous peptide microarray

• US 9,977,019 B2
• Filed: 04/16/2013
• Issued: 05/22/2018
• Est. Priority Date: 04/25/2012
• Status: Active Grant

• Current Assignee: Ms Array LLC
• Original Assignee: Ms Array LLC
• Inventors: Yu, Marvin
• Primary Examiner(s): LEAVITT, MARIA GOMEZ
• Application Number:
  US14/394,435
• Publication Number:
  US 20150192578A1
• Time in Patent Office: 1,862 Days
• Field of Search: None
• CPC Class Codes: G01N 2333/90 Enzymes; Proenzymes
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

A fluorous peptide microarray, a process of detecting information by using a fluorous peptide microarray, and a process of forming a fluorous peptide microarray are disclosed. The fluorous peptide microarray includes a covalently-modified conductive fluorous surface and fluorous-tagged peptides having natural amino acids positioned on the covalently-modified conductive fluorous surface. The fluorous-tagged peptides are configured for analysis of one or both of a protein and an enzyme. The process of detecting information includes using the fluorous peptide microarray. The process of forming the fluorous peptide microarray includes spotting the fluorous-modified composition to the fluorous surface after a solution phase reaction or transfer blotting the fluorous-modified composition.

Claims

1. A microarray for detecting targets and biomolecule interactions, comprising:

(a) a covalently-modified electrically conductive fluorous surface comprising a compositional grid,

(i) wherein the surface is electrically conductive with a sheet resistivity of less than 50 ohms/square, and

(ii) wherein the electrically conductive surface is a perfluorocarbon modified;

(b) a plurality of fluorous-modified peptide compositions, wherein each fluorous-modified peptide composition in the plurality of fluorous-modified peptide compositions includes;

(i) a terminal fluorous domain or tag comprising at least one perfluoroalkyl groups in the fluorous domain or tag, wherein the terminal fluorous domain or tag immobilizes each fluorous-modified peptide composition on the perfluorocarbon covalently-modified electrically conductive surface by non-covalently interacting with the perfluorocarbon modified surface through fluorous partitioning, wherein each fluorous-modified peptide composition individually occupies a defined location within the grid,

(ii) a diamine linker connected to the terminal fluorous domain or tag; and

(iii) a peptide connected to the diamine linker, wherein the peptide is configured to bind targets and interact with biomolecules of interest;

(c) wherein the presence of a target bound to the peptide in the fluorous-modified peptide composition or biomolecule interaction with the peptide in the fluorous-modified peptide composition is detectable by mass spectrometry through individual laser ionization of the peptide fluorous-modified composition immobilized on the perfluorocarbon covalently-modified electrically conductive surface.

2. The microarray of claim 1, wherein the perfluorocarbon modified surface includes a fluorous-modified indium tin oxide, a fluorous-modified silicon black, or a combination thereof.

3. The microarray of claim 1, wherein the peptide in each fluorous-modified peptide composition is adapted to undergo enzyme modification.

4. The microarray of claim 1, wherein the peptide in each fluorous modified peptide composition is configured to interact with kinases, proteases, glycosyl transferases, phosphatases, histone modifying enzymes, or a combination thereof.

5. The microarray of claim 1, wherein the peptide in each fluorous modified peptide composition is an irreversible binding partner for interacting with proteins or protein complexes.

6. The microarray of claim 1, wherein the peptide in each fluorous modified peptide composition is a reversible binding partner for interacting with proteins or protein complexes.

7. The microarray of claim 1, wherein the mass spectrometry further includes matrix assistance analysis methods, matrix-free analysis methods, or a combination thereof.

8. The microarray of claim 1, wherein the microarray detects a change in mass of a fluorous-modified composition through action of an enzyme by mass spectrometry.

9. The microarray of claim 1, wherein the microarray detects a change in mass of a fluorous-modified composition through binding of a protein by mass spectrometry.

10. The microarray of claim 1, wherein the targets include proteins and enzymes, and wherein the biomolecule interactions include enzymatic modification of the peptides in the fluorous-modified compositions.

11. The microarray of claim 1, wherein the peptide in each fluorous-modified peptide compositions interact with a target or biomolecule after the immobilization of the fluorous-modified compositions on the perfluorocarbon modified electrically conductive surface.

12. The microarray of claim 1, wherein the peptide in each fluorous-modified peptide compositions interact with a target or biomolecule prior to the immobilization of the fluorous-modified compositions on the perfluorocarbon modified electrically conductive surface.

13. The microarray of claim 1, wherein the perfluorocarbon modified electrically conductive surface is washed prior to mass spectrometry to selectively retain the fluorous-modified compositions having bound targets or biomolecule interactions.

14. The microarray of claim 1, wherein the microarray is used to detect information, and wherein the information includes the presence of bound targets, the presence of biomolecule interactions, or a combination thereof.

15. The microarray of claim 1, further comprising a solid-phase attachment group connected to the linker within the fluorous-modified compositions.